Marine Sponge Haliclona Research Articles

Papuamine and haliclonadiamine, obtained from an Indonesian sponge Haliclona sp., inhibited cell proliferation of human cancer cell lines

The extract of an Indonesian marine sponge Haliclona sp. showed potent cytotoxicity against human solid cancer cell lines, MCF-7 (breast), LNCap (prostate), Caco-2 (colon) and HCT-15 (colon) cells. Study on nuclear morphological changes and flow cytometric analysis suggested that the component(s) in the extract would induce an apoptosis to these cancer cells. Bioassay-guided isolation yielded two pentacyclic alkaloids, papuamine (1) and haliclonadiamine (2), which inhibited cell proliferation of six human cancer cell lines with IC50 values of 0.93–1.50 and 1.00–4.44 µM, respectively. Compounds 1 and 2 accumulated lymphoma U937 cells at sub-G1 phase and induced a condensation of chromatin and fragmentation of nucleus.

Antifilarial activity of marine sponge Haliclona oculata against experimental Brugia malayi infection

The present study incorporates the findings on in vitro and in vivo antifilarial activity in the marine sponge, Haliclona oculata using an experimental rodent infection of human lymphatic filarial parasite, Brugia malayi. The in vitro antifilarial action was determined on both adult female worms as well as microfilariae using two parameters viz. adverse effect on motility and inhibition in MTT reduction by the treated adult parasite over control worm. The antifilarial activity could be located in the methanol extract and one of its four fractions (chloroform). Bioactivity guided fractionation of chloroform fraction led to localization of in vitro activity in one of its eight chromatographic fractions. Methanol extract, chloroform fraction and one of the chromatographic fractions revealed IC50 values of 5.00, 1.80, and 1.62μg/ml, respectively when adult B. malayi were exposed to these test samples for 72h at 37°C. Under similar exposure conditions, the IC50 values for microfilariae were 1.88, 1.72 and 1.19μg/ml, respectively. The active test samples were found to be safe revealing >10 selectivity indices (SI) on the basis of cytotoxicity to Vero cells (monkey kidney cells) and therefore selected for in vivo evaluation against primary (adult B. malayi intraperitoneal transplanted jird) and secondary (subcutaneous infective larvae induced mastomys) screens. In primary jird model, the three test samples at 100mg/kg for five consecutive days by subcutaneous route demonstrated significant macrofilaricidal efficacy to the tune of 51.3%, 64% and 70.7% by methanol extract, chloroform and chromatographic fraction, respectively. The three samples demonstrated 45–50% macrofilaricidal activity with moderate embryostatic effect in secondary model at 5×500, 5×250 and 5×125mg/kg by oral route. Chromatographic fraction possessing highest antifilarial action was primarily found to be a mixture of four alkaloids Mimosamycin, Xestospongin-C, Xestospongin-D and Araguspongin-C in addition to few minor compounds.

In vitro antiplasmodial activity of bacterium RJAUTHB 14 associated with marine sponge Haliclona Grant against Plasmodium falciparum

Malaria is the most important parasitic disease, leading to annual death of about one million people, and the Plasmodium falciparum develops resistance to well-established antimalarial drugs. The newest antiplasmodial drug from a marine microorganism helps in addressing this problem. In the present study, Haliclona Grant were collected and subjected for enumeration and isolation of associated bacteria. The count of bacterial isolates was maximum in November 2007 (18 × 10(4) colony-forming units (CFU) g(-1), and the average count was maximum during the monsoon season (117 × 10(3) CFU g(-1)). Thirty-three morphologically different bacterial isolates were isolated from Haliclona Grant, and the extracellular ethyl acetate extracts were screened for antiplasmodial activity against P. falciparum. The antiplasmodial activity of bacterium RJAUTHB 14 (11.98 μg[Symbol: see text]ml(-1)) is highly comparable with the positive control chloroquine (IC(50) 19.59 μg[Symbol: see text]ml(-1)), but the other 21 bacterial extracts showed an IC(50) value of more than 100 μg[Symbol: see text]ml(-1). Statistical analysis reveals that significant in vitro antiplasmodial activity (P < 0.05) was observed between the concentrations and time of exposure. The chemical injury to erythrocytes showed no morphological changes in erythrocytes by the ethyl acetate extract of bacterial isolates after 48 h of incubation. The in vitro antiplasmodial activity might be due to the presence of reducing sugars and alkaloids in the ethyl acetate extracts of bacterium RJAUTHB 14. The 16S rRNA gene partial sequence of bacterium RJAUTHB 14 is deposited in NCBI (GenBank accession no. GU269569). It is concluded from the present study that the ethyl acetate extracts of bacterium RJAUTHB 14 possess lead compounds for the development of antiplasmodial drugs.

Diversity and bioactive potential of endospore-forming bacteria cultured from the marine sponge Haliclona simulans

Despite the frequent isolation of endospore-formers from marine sponges, little is known about the diversity and characterization of individual isolates. The main aims of this study were to isolate and characterize the spore-forming bacteria from the marine sponge Haliclona simulans and to examine their potential as a source for bioactive compounds. A bank of presumptive aerobic spore-forming bacteria was isolated from the marine sponge H. simulans. These represented c. 1% of the total culturable bacterial population. A subgroup of thirty isolates was characterized using morphological, phenotypical and phylogenetic analysis. A large diversity of endospore-forming bacteria was present, with the thirty isolates being distributed through a variety of Bacillus and Paenibacillus species. These included ubiquitous species, such as B. subtilis, B. pumilus, B. licheniformis and B. cereus group, as well as species that are typically associated with marine habitats, such as B. aquimaris, B. algicola and B. hwajinpoensis. Two strains carried the aiiA gene that encodes a lactonase known to be able to disrupt quorum-sensing mechanisms, and various isolates demonstrated protease activity and antimicrobial activity against different pathogenic indicator strains, including Clostridium perfringens, Bacillus cereus and Listeria monocytogenes. The marine sponge H. simulans harbours a diverse collection of endospore-forming bacteria, which produce proteases and antibiotics. This diversity appears to be overlooked by culture-dependent and culture-independent methods that do not specifically target sporeformers. Marine sponges are an as yet largely untapped and poorly understood source of endospore-forming bacterial diversity with potential biotechnological, biopharmaceutical and probiotic applications. These results also indicate the importance of combining different methodologies for the comprehensive characterization of complex microbial populations such as those found in marine sponges.

Multiple Approaches To Enhance the Cultivability of Bacteria Associated with the Marine Sponge Haliclona ( gellius ) sp

Three methods were examined to cultivate bacteria associated with the marine sponge Haliclona (gellius) sp.: agar plate cultures, liquid cultures, and floating filter cultures. A variety of oligotrophic media were employed, including media with aqueous and organic sponge extracts, bacterial signal molecules, and siderophores. More than 3,900 isolates were analyzed, and 205 operational taxonomic units (OTUs) were identified. Media containing low concentrations of mucin or a mixture of peptone and starch were most successful for the isolation of diversity, while the commonly used marine broth did not result in a high diversity among isolates. The addition of antibiotics generally led to a reduced diversity on plates but yielded different bacteria than other media. In addition, diversity patterns of isolates from agar plates, liquid cultures, and floating filters were significantly different. Almost 89% of all isolates were Alphaproteobacteria; however, members of phyla that are less commonly encountered in cultivation studies, such as Planctomycetes, Verrucomicrobia, and Deltaproteobacteria, were isolated as well. The sponge-associated bacteria were categorized into three different groups. The first group represented OTUs that were also obtained in a clone library from previously analyzed sponge tissue (group 1). Furthermore, we distinguished OTUs that were obtained from sponge tissue (in a previous study) but not from sponge isolates (group 2), and there were also OTUs that were not obtained from sponge tissue but were obtained from sponge isolates (group 3). The 17 OTUs categorized into group 1 represented 10 to 14% of all bacterial OTUs that were present in a large clone library previously generated from Haliclona (gellius) sp. sponge tissue, which is higher than previously reported cultivability scores for sponge-associated bacteria. Six of these 17 OTUs were not obtained from agar plates, which underlines that the use of multiple cultivation methods is worthwhile to increase the diversity of the cultivable microorganisms from sponges.

Antifungal activity of marine sponge Haliclona exigua (Krikpatrick)

Summary The methanol extract of the marine sponge Haliclona exigua showed promising antifungal activity in vitro models against Candida albicans (MIC = 7.8 μg/ml), Cryptococcus neoformans (MIC = 31.2 μg/ml), Sporothrix schenckii (MIC = 31.2 μg/ml), Trichophyton mentagrophytes (MIC = 31.2 μg/ml), Aspergillus fumigatus (MIC = 31.2 μg/ml) and Candida parapsilosis (MIC = 7.8 μg/ml). After division of the methanolic extract into four fractions, the antifungal activity was detected exclusively in the hexane-soluble and the chloroform-soluble. Both fractions were combined and the major active compounds of these extracts were purified. The purified compound, araguspongin C, showed promising results against Cryptococcus neoformans , Sporothrix schenckii , Trichophyton mentagrophytes and Aspergillus fumigatus with identical MIC of 50 μg/ml against these fungi.

Streptomyces tateyamensis sp. nov., Streptomyces marinus sp. nov. and Streptomyces haliclonae sp. nov., isolated from the marine sponge Haliclona sp.

Three Gram-positive, NaCl-requiring actinobacteria were isolated from a marine sponge, Haliclona sp., collected from the coast of Tateyama City, Japan. Comparison of 16S rRNA gene sequences indicated that these strains represent novel members of the genus Streptomyces, exhibiting low 16S rRNA gene sequence similarities of 98.3-97.4 % with recognized members of the genus. The cell hydrolysates contained the LL-isomer of diaminopimelic acid and the predominant quinones were MK-9 (H(6) and/or H(8)). The DNA G+C contents were in the range 72-75mol%. A polyphasic study of the strains and comparison of the characters with related species of the genus show that these strains represent three novel species of the genus Streptomyces. Therefore, the names Streptomyces tateyamensis sp. nov., Streptomyces haliclonae sp. nov. and Streptomyces marinus sp. nov. are proposed for strains Sp080513SC-30(T) (=NBRC 105048(T) =DSM 41969(T)), Sp080513SC-31(T) (=NBRC 105049(T) =DSM 41970(T)) and Sp080513GE-26(T) (=NBRC 105047(T) =DSM 41968(T)), respectively.

Endoglucanase activities and growth of marine-derived fungi isolated from the spongeHaliclona simulans

The conversion of cheap cellulosic biomass to more easily fermentable sugars requires the use of costly cellulases. We have isolated a series of marine sponge-derived fungi and screened these for cellulolytic activity to determine the potential of this unique environmental niche as a source of novel cellulase activities. Fungi were isolated from the marine sponge Haliclona simulans. Phylogenetic analysis of these and other fungi previously isolated from H. simulans showed fungi from three phyla with very few duplicate species. Cellulase activities were determined using plate-based assays using different media and sea water concentrations while extracellular cellulase activities were determined using 3,5-dinitrosalicylic acid (DNSA)-based assays. Total and specific cellulase activities were determined using a range of incubation temperatures and compared to those for the cellulase overproducing mutant Hypocrea jecorina QM9414. Several of the strains assayed produced total or relative endoglucanase activities that were higher than H. jecorina, particularly at lower reaction temperatures. Marine sponges harbour diverse fungal species and these fungi are a good source of endoglucanase activities. Analysis of the extracellular endoglucanase activities revealed that some of the marine-derived fungi produced high endoglucanase activities that were especially active at lower temperatures. Marine-derived fungi associated with coastal marine sponges are a novel source of highly active endoglucanases with significant activity at low temperatures and could be a source of novel cellulase activities.

Comparison of two marine sponge-associatedPenicillium strains DQ25 and SC10: differences in secondary metabolites and their bioactivities

Two strains ofPenicillium, DQ25 and SC10, isolated from marine spongeHaliclona angulata (Bowerbank) andHymeniacidon sp. respectively, were subjected to stationary cultivation under GYP medium for 30 days. The fermentation extracts were undergone bioactivities assays against human pathogens, phytopathogenic fungi and brine shrimp (Artemia salina). Bioassays-guided compounds isolation was performed by Silica gel columns and Sephadex LH-20 chromatography. Spectroscopic methods were used to structures elucidation of the compounds. Results showed the activities of secondary metabolites of strain DQ25 were generally stronger than that of strain SC10. Major bioactive molecules isolated from strain DQ25 were a 1,4-naphthoquinone derivative and an unidentified alkaloid. The two components were not isolated from the extract of strain SC10. ITS sequences revealed that these two species have the greatest similarity withPenicillium vinaceum andPenicillium granulatum respectively.

In vitro and in vivo antifilarial potential of marine sponge, Haliclona exigua (Kirkpatrick), against human lymphatic filarial parasite Brugia malayi

The present study reports on the antifilarial activity of a marine sponge Haliclona exigua (phylum Porifera). The crude methanol extract and n-butanol-soluble fraction killed adult Brugia malayi at 31.25-microg/ml concentration (both in motility and 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide assay) while the chloroform fraction was lethal at a lower concentration of 15.6 microg/ml. The activity could be located in a single molecule araguspongin C which brought about mortality of worm at 15.6 microg/ml. In vivo evaluation of the crude extract (5 x 500 mg/kg, orally) and the chloroform fraction (5 x 250 mg/kg, orally) in B. malayi-infected rodent host, Mastomys coucha, did not show any significant microfilaricidal actions; however, microfilarial densities in both the treated groups were significantly much lower than those of untreated group in contrast to standard filaricide diethylcarbamazine which exerted 79% microfilaricidal action on day 8 of treatment. Both these extracts also demonstrated adulticidal (macrofilaricidal) activity which was more pronounced in the chloroform fraction (50.2%). In addition, there was moderate adverse effect on the reproductive potential of female worms (crude extract 46.5%; chloroform 58.6%). The findings suggest that the marine sponge H. exigua possesses adulticidal and embryostatic action against human lymphatic filarial parasite B. malayi in experimental rodent model and this activity could be attributed to the presence of araguspongin C.

Nucleosides from the Marine Sponge Haliclona sp.

Three known nucleosides were isolated from the sponge Haliclona sp. The structures were established on the basis of NMR data and comparison with those reported, and chemotaxonomic relationships of the sponge nucleosides were discussed.

Haliclonin A, a New Macrocyclic Diamide from the Sponge Haliclona sp.

Haliclonin A (1), a macrocyclic diamide of a novel skeletal class, was isolated from the marine sponge Haliclona sp. collected from Korean waters. The structure of this compound was determined using a combination of spectroscopic and chemical analyses. The new compound exhibited moderate cytotoxicity and antibacterial activity against diverse microbial strains.

Isolation and structure determination of the biologically active sphingolipids from marine sponge Haliclona species

In a continuation to our study on the marine sponge Haliclona species we have isolated three new cytotoxic components of sphingolipids (1–3). Methanolysis of the sphingolipid 1a–d in methanol produces fatty acid methyl ester. GC/MS was used to determine the length. The structure of each isolated compound has been determined on the basis of spectroscopic data and chemical evidence.

New Cerebrosides from a Marine Sponge Haliclona (Reniera) sp.

A chemical investigation of the MeOH extract of a marine sponge Haliclona (Reniera) sp., collected off the coast of Ulleung Island, Korea, led to the isolation of thirteen new cerebrosides (1--3, 5--14), along with a known analogue (4). Their structures were elucidated on the basis of 1D and 2D NMR spectroscopy, MS spectrometry, and chemical method. The major new features of these glucocerebrosides are C15 and C19 acyl chains, long (C24-C28) acyl chains, or the S-configuration of the acyl chains. It is noteworthy that both R- and S-configurations of the acyl chains were observed in the same specimen.

2-Palmitamidoethanesulfonic acid, a taurine derivative from the marine sponge Haliclona sp.

[Wang, Bin; Zhang, Si; Liu, Yonghong] Chinese Acad Sci, Key Lab Marine Bioresources Sustainable Utilizat, S China Sea Inst Oceanol, Guangzhou 510301, Guangdong, Peoples R China; [Wang, Bin] Beihua Univ, Jilin 132001, Peoples R China; [Lee, Kyung Jin] Hannam Univ, Dept Biol, Taejon, South Korea; [Jung, Jee H.] Pusan Natl Univ, Coll Pharm, Pusan 609735, South Korea

Isolation and Analysis of Bacteria with Antimicrobial Activities from the Marine Sponge Haliclona simulans Collected from Irish Waters

Samples of the marine sponge Haliclona simulans were collected from Irish coastal waters, and bacteria were isolated from these samples. Phylogenetic analyses of the cultured isolates showed that four different bacterial phyla were represented; Bacteriodetes, Actinobacteria, Proteobacteria, and Firmicutes. The sponge bacterial isolates were assayed for the production of antimicrobial substances, and biological activities against Gram-positive and Gram-negative bacteria and fungi were demonstrated, with 50% of isolates showing antimicrobial activity against at least one of the test strains. Further testing showed that the antimicrobial activities extended to the important pathogens Pseudomonas aeruginosa, Clostridium difficile, multi-drug-resistant Staphylococcus aureus, and pathogenic yeast strains. The Actinomycetes were numerically the most abundant producers of antimicrobial activities, although activities were also noted from Bacilli and Pseudovibrio isolates. Surveys for the presence of potential antibiotic encoding polyketide synthase and nonribosomal peptide synthetase genes also revealed that genes for the biosynthesis of these secondary metabolites were present in most bacterial phyla but were particularly prevalent among the Actinobacteria and Proteobacteria. This study demonstrates that the culturable fraction of bacteria from the sponge H. simulans is diverse and appears to possess much potential as a source for the discovery of new medically relevant biological active agents.

Spongiibacter marinus gen. nov., sp. nov., a halophilic marine bacterium isolated from the boreal sponge Haliclona sp. 1

Strain HAL40b(T) was isolated from the marine sponge Haliclona sp. 1 collected at the Sula Ridge off the Norwegian coast and characterized by physiological, biochemical and phylogenetic analyses. The isolate was a small rod with a polar flagellum. It was aerobic, Gram-negative and oxidase- and catalase-positive. Optimal growth was observed at 20-30 degrees C, pH 7-9 and in 3 % NaCl. Substrate utilization tests were positive for arabinose, Tween 40 and Tween 80. Enzyme tests were positive for alkaline phosphatase, esterase lipase (C8), leucine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and N-acetyl-beta-glucosaminidase. The predominant cellular fatty acid was C(17 : 1) omega8, followed by C(17 : 0) and C(18 : 1) omega7. Analysis by matrix-assisted laser desorption/ionization time-of-flight MS was used to characterize the strain, producing a characteristic low-molecular-mass protein pattern that could be used as a fingerprint for identification of members of this species. The DNA G+C content was 69.1 mol%. Phylogenetic analysis supported by 16S rRNA gene sequence comparison classified the strain as a member of the class Gammaproteobacteria. Strain HAL40b(T) was only distantly related to other marine bacteria including Neptunomonas naphthovorans and Marinobacter daepoensis (type strain sequence similarity >90 %). Based on its phenotypic, physiological and phylogenetic characteristics, it is proposed that the strain should be placed into a new genus as a representative of a novel species, Spongiibacter marinus gen. nov., sp. nov.; the type strain of Spongiibacter marinus is HAL40b(T) (=DSM 17750(T) =CCUG 54896(T)).

Renierosides, Cerebrosides from a Marine Sponge Haliclona (Reniera) sp.

Guided by the brine shrimp lethality assay, eight new cerebrosides (1-8) have been isolated from an extract of the marine sponge Haliclona (Reniera) sp. A novel feature of these cerebrosides was the presence of unprecedented amide-linked long-chain fatty acid moieties. The planar structures of the cerebrosides (1-8) were established by 1D and 2D NMR spectroscopic techniques, mass spectrometric analyses, and chemical degradation methods. The isolated compounds did not display cytotoxicity to a panel of five human solid tumor cell lines.

The marine sponge Haliclona simulans harbours a novel and diverse microbial community with extensive genetic diversity in polyketide synthase genes, with potential biopharmaceutical applications

The marine sponge Haliclona simulans harbours a novel and diverse microbial community with extensive genetic diversity in polyketide synthase genes, with potential biopharmaceutical applications

Temperature and Spatiotemporal Variability of Salicylihalamide A in the Sponge Haliclona sp.

The production of salicylihalamide A by the marine sponge Haliclona sp. was investigated. Samples of the two morphologies (green and brown) were collected from four locations covering approximately 1,200 km of coastline. Temporal variation between winter and summer was also examined at Bremer Bay. Chemical profiling by using liquid chromatography coupled with ultra violet detection and mass spectrometry showed that salicylihalamide A was produced only by the green morphology. Salicylihalamide A concentration was significantly correlated to water temperature but not to the size or depth of the sponge. Salicylihalamide A concentration was found to differ significantly among locations (Bremer Bay 13.5 microg g(-1), Hamelin Bay 11 microg g(-1), Rottnest Island 9.9 microg g(-1), and Jurien Bay 8.5 microg g(-1)) partially accounted for by the influence of water temperature. A difference between seasons was also observed in Bremer Bay (summer concentration of 13.5 microg g(-1) vs. winter concentration of 8.2 microg g(-1)). Environmental and physiological factors appear to be important in the production of salicylihalamide A by the green morphology. Additionally, the brown morphology does not produce salicylihalamide A, thus adding to the evidence that this morphology may be a different species.